Use of Alternan as Texturizing Agent in Foodstuffs and Cosmetics

ABSTRACT

The invention is directed to the use of alternan as texturizing agent, particularly as fat or oil replacer in foodstuffs or cosmetic preparations, a homogeneous composition comprising alternan and water, the use of the homogeneous composition as texturizing agent in foodstuffs or cosmetic preparations, and foodstuffs and cosmetic preparations comprising alternan as texturizing agent or a homogeneous composition comprising alternan.

The present invention is directed to the use of alternan as texturizing agent, particularly as fat or oil replacer in foodstuffs or cosmetic preparations, a homogeneous composition comprising alternan and water, the use of the homogeneous composition as texturizing agent in foodstuffs or cosmetic preparations, and foodstuffs and cosmetic preparations comprising alternan as texturizing agent or a homogeneous composition comprising alternan.

In recent years, problems of obesity and hyperlipemia have increased due to excessive consumption of oils, fats, sugars, etc., and decreased consumption of dietary fibers. It has been suggested that certain adult diseases, which are leading causes of death, such as cancer, heart diseases, etc., are associated with excessive consumption of oils, fats, and sugars and decreased consumption of dietary fibers. For these reasons, low calorie foodstuffs wherein oil or fat is replaced by substances with beneficial texturizing properties have become popular.

Some Polysaccharides have been described as texturizing agents and fat replacers. U.S. Pat. No. 5,169,671 relates to a food or drink containing a fructose polymer mainly composed of beta-2,1-bond, referred to as “polyfructan”, as a substitute for oils, fats, and sugars and having improved gel properties compared with foods containing other oil, fat, or sugar substitutes and a creamy taste and texture equivalent to that derived from oils and fats and a method of producing such a food or drink.

WO2007/128559 and WO2007/128559 teach that very long chain inulins are efficient texturizing agents and fat replacers, especially in dairy products and ice cream.

However, in view of the increased need for efficient texturizing agents and fat replacers for foods it was an object of the present invention to find alternative substances which can be used for this purpose

The present invention is directed to the use of alternan as a texturizing agent in foodstuffs or cosmetic preparations.

In another aspect the present invention is directed to the use of alteman as an emulsifier, preferably for foodstuffs and cosmetic preparations.

A texturizing agent according to the present invention imparts texture to foodstuffs or cosmetic preparations or modifies the texture of foodstuffs or cosmetic preparations. The term “texture” refers to the properties held and sensations caused by the external surface of foodstuffs or cosmetic preparations received through the sense of touch.

In connection with foodstuffs, the term “mouthfeel” is used alternatively to the term “texture” and both terms are used herein interchangeably with respect to foodstuffs. In connection with cosmetic preparations, a major meaning of the term texture is the skin feel of a cosmetic preparation.

In the present invention it can be shown that alternan is a very suitable fat or oil replacer in foodstuffs. The fat or oil component of foodstuffs has usually a major impact on its texture/mouthfeel, organoleptic characteristics, and flavor. A fat or oil content usually imparts a creamy and smooth mouthfeel to a foodstuff. These desired properties of foodstuffs can also be reached by adding alteman to foodstuffs. The term “fat or oil replacer” in this connection means that at least a part of a fat or oil component in a foodstuff is removed from the foodstuff and replaced by alternan. The term “fat or oil replacer”, however, can also mean that the total fat or oil component in a foodstuff is removed from the foodstuff and replaced by alternan.

Alternan is a polysaccharide composed of glucose units. The glucose units are linked to each other via a-1,3- and a-1,6-glycosidic bonds, and said two types of bonds predominantly appear alternatingly. Alternan may contain branches (Seymour et al., Carbohydrate Research 74, (1979), 41-62). Alternan and methods for producing alternan are well known from the state of the art. See for example Jeanes et al. (1954) J. Am. Chem. Soc., 76: 5041-5052, Misaki et al. (1980) Carbohydr. Res., 84: 273-285, Cote and Robyt (1982), Carbohydr. Res., 101: 57-74, Cote (1992), Carbohydrate Polymers 19, 249-252, WO 00/47727, U.S. Pat. No. 5,702,942, US20060127328, PCT/EP2008/051760.

Alternan according to the present invention preferably has a weight average molecular weight Mw in the range of 10 000 000 g/mol to 60 000 000 g/mol (determined with GPC MALLS), more preferably in the range of 12 000 000 g/mol to 50 000 000 g/mol. In a special embodiment, alteman is produced with alternansucrase originating from Leuconostoc Mesenteroides as described in WO 00/47727 and shows a weight average molecular weight Mw in the range of 33 000 000 g/mol to 60 000 000 g/mol (determined with GPC MALLS), more preferably in the range of 33 000 000 g/mol to 50 000 000 g/mol. In still another special embodiment, alternan is produced with truncated alternansucrase enzyme as described in PCT/EP2008/051760 and shows a weight average molecular weight Mw in the range of 12 000 000 g/mol to 30 000 000 g/mol (GPC MALLS), more preferably in the range of 14 000 000 g/mol to 28 000 000 g/mol, still more preferably in the range of 16 000 000 g/mol to 26 000 000 g/mol, most preferably 19 000 000 g/mol to 23 000 000 g/mol. Truncated alternansucrase enzymes, methods for producing alternan therefrom as well as the alternan itself are described in PCT/EP2008/051760, which is incorporated herein by reference in its entirety.

A preferred foodstuff wherein alteman is used as a fat replacer is selected from dairy products, yoghurts, ice creams, milk-based soft ice, milk-based garnishes, puddings, cream, whipped cream, chocolate cream, butter cream, creme fraiche, curd, milk, such as skim milk, buttermilk, soured milk, kefir, milkshakes, egg custard, cheese, such as cream cheese, soft cheese, sliced cheese, hard cheese, nutrition bars, energy bars, breakfast bars, confectionery, bakery products, crackers, cookies, biscuits, cereal chips, snack products, diet drinks, finished drinks, sports drinks, stamina drinks, powdered drink mixtures for dietary supplementation, infant and baby foodstuff, bread, croissants, breakfast cereals, spreads, sugar-free biscuits and chocolates, calcium chews, meat products, sausages, mayonnaise, dressings, nut butter, deep-frozen meals, sauces, gravy, soups, shortenings, canned foods and ready-to-serve meals.

The level of alternan in the foodstuff depends on the kind of foodstuff. Usually, alteran is used in an amount which is sufficient to reach the desired degree of fat/oil replacement and the desired mouthfeel properties. Without limitation exemplary levels of alteman in foodstuffs are 0.1-10 weight percent, more preferably 0.1-5 weight percent, and most preferably 0.1-3 weight percent, based on the total weight of all components forming the foodstuff.

Oils and fats may be wholly substituted with alternan. However, a preferred degree of substitution is 20 to 80% by weight based on weight of the oils and/or fats.

In a further aspect the present invention is directed to a foodstuff comprising alternan as texturizing agent, particularly as a fat or oil replacer. The foodstuff is preferably selected from the above mentioned foodstuffs The term “foodstuff” according to the invention also encompasses beverages.

The present invention is also directed to a cosmetic preparation, comprising alternan as texturizing agent. The cosmetic preparation where alternan may be employed include, but are not limited to, deodorants, antiperspirants, antiperspirant/deodorants, shaving products, skin creams and lotions, moisturizers, toners, bath products, cleansing products, hair care products such as shampoos, conditioners, mousses, styling gels, nails creams and lotions, cuticle softeners, protective creams such as sunscreen, anti-aging products, and other personal care formulations.

In a further aspect, the present invention is directed to a homogeneous composition comprising alternan and water. The term “homogeneous composition” means that the composition shows no visible phase separation or separation of its constituents (not visible to the naked eye).

Depending on the relative amounts of alternan and water, said homogeneous composition can show a wide viscosity range. So, the homogeneous composition may be a liquid or a more viscous cream, wherein a cream is preferred.

In one embodiment the composition according to the invention comprises 5-15 weight-% alternan and 85-95 weight-% water, based on the total weight of the composition, more preferably 8-13 weight-% alternan and 87-92 weight-% water. Further components may be included, such as surfactants and salts.

In another embodiment, a homogeneous composition according to the invention comprises alternan, water and at least one fat or oil or a mixture of fat(s) and oil(s). Said homogeneous composition comprising alternan, water and at least one fat and/or oil shows no visible phase separation between the fat or oil component and the water component (not visible to the naked eye). Thus, said composition comprising alternan, water and at least one fat and/or oil may also be designated as an emulsion.

Depending on the relative amounts of alternan, water, and oil and/or fat, said homogeneous composition can show a wide viscosity range. So, the homogeneous composition may be a liquid or a more viscous cream, wherein a cream is preferred.

In one embodiment a composition comprises 5-15 weight-% alternan, 70-90 weight-% water, 5-15 weight-% fat or oil, more preferably 8-13 weight-% alternan, 74-84 weight-% water, and 8-13 weight-% fat or oil, based on the total weight of the composition. Further components such as surfactants and salts may be included.

Preferred fats/oils as a component for a homogeneous composition comprising alternan, water and at least one fat and/or oil are selected from arachidic oil, avocado oil, cotton seed oil, safflower oil, peanut oil, hazelnut oil, hemp oil, jojobaoil, camenillia oil, cocoa butter, coconut oil, pumpkin oil, linseed oil, macadamia nut oil, corn germ oil, almond oil, mango seed fat, poppy seed oil, evening primrose oil, olive oil, palm oil, palm kernel oil, papaya oil, pistachio nut oil, pecan nut oil, rapeseed oil, castor oil, mustard seed oil, sesame oil, rhea butter, soybean oil, sunflower oil, walnut oil, water melonseed oil, grapeseed oil, wheat germ oil, cedarwood oil, wherein said oils might optionally be hardened oils.

In still another aspect, the present invention is directed to a homogeneous composition comprising alternan and milk. In a preferred embodiment said composition comprises 5-15 weight-% alternan and 85-95 weight-% milk, based on the total weight of the composition, more preferably 8-13 weight-% alternan and 87-92 weight-% milk. Further components may be included, such as surfactants and salts. A cream comprising alternan and milk might preferably be used for all kind of dairy products.

The viscosity of a homogeneous composition according to the invention is adapted to the desired kind of foodstuff or cosmetic preparation, i.e. to the desired final viscosity of said foodstuff or cosmetic. However, as said above, the homogeneous composition is preferably a cream, wherein a preferred range of viscosity of a cream is from 0.5 to 1.5 Pa*s, more preferably 0.8 to 1.3 Pa*s at 20° C. and a shear rate of 40 s⁻¹.

All of the above described homogeneous compositions, preferably creams, are hereinafter also designated as “(homogeneous) composition according to the invention” or “cream according to the invention”.

The present invention also relates to a method for preparing the above described compositions, the method comprising

-   -   a) giving alternan and water into a vessel     -   b) applying shear to obtain a homogeneous composition, and         optionally     -   c) adding oil or fat and applying shear to obtain a homogeneous         composition.

The optional step c) relates to the above described embodiment of a homogeneous composition, comprising alteman, water and oil or fat.

Mixing in steps b) and c) is preferably done under high shear with commonly known devices for such purpose, such as Ultra Turrax® dispersing tool from IKA company. In a preferred embodiment, steps a) and b) are performed simultaneously, e.g. by adding alteman as a powder to water with simultaneous application of high shear. Optional components as salts and emulsifiers might be added at any stage of the preparation.

In a further aspect, the present invention is related to the use of a homogeneous composition as described above for the manufacture of foodstuffs or cosmetic preparations. In this connection, said homogeneous compositions, preferably creams, can preferably used for above-listed foodstuffs and cosmetics.

According to the present invention the homogeneous compositions can be used as constituent for foodstuffs or cosmetic preparations. In this connection, homogeneous compositions according to the present invention can fulfill, without limitation, following functions: giving body to foodstuffs or cosmetic preparations, act as a base for foodstuffs or cosmetic preparations, act as a texturizing agent for foodstuffs or cosmetic preparations. The homogeneous compositions according to the present invention can fulfill one or more of the aforementioned functions.

In regard to texturizing properties the homogeneous compositions according to the present invention are preferably used as a fat or oil replacer in foodstuffs. In this connection, the homogeneous composition is preferably a cream. The preferred foodstuffs for said use are selected from dairy products, yoghurts, ice creams, milk-based soft ice, milk-based garnishes, puddings, cream, whipped cream, chocolate cream, butter cream, creme fraiche, curd, milkshakes, egg custard, cheese, such as cream cheese, soft cheese, sliced cheese, hard cheese, nutrition bars, energy bars, breakfast bars, confectionery, bakery products, crackers, cookies, biscuits, cereal chips, snack products, infant and baby foodstuff, bread, croissants, spreads, sugar-free biscuits and chocolates, calcium chews, meat products, sausages, mayonnaise, dressings, nut butter, deep-frozen meals, sauces, gravy, soups, shortenings, canned foods and ready-to-serve meals.

Homogeneous compositions according to the invention have a texture similar to oils or fats. By substituting this composition for liquid oils and solid fats, for example in dressings, mayonnaise, fresh cream, cream cheese, butter, salad oil, etc., in various cooked and processed food, the number of calories in the resulting food are greatly reduced. Oils and fats may be wholly substituted with a homogeneous composition according to the present invention. However, a preferred degree of substitution is 20 to 80% by weight based on the oils and fats.

Finally, the present invention is also directed to a foodstuff or cosmetic preparation, characterized in that during its manufacturing process a homogeneous composition according to the present invention was added as a constituent. In the foodstuff or cosmetic, a homogeneous composition according to the present invention preferably fulfills the functions as described above with respect to its use. Depending on the kind of foodstuff or cosmetic and its manufacturing process, the homogeneous composition might be present in the final product without substantial structural alteration. This might for example be the case when a cream of the invention is used as the base for a cosmetic preparation and only some active ingredients are added. In such case, the final foodstuff or cosmetic preparation is characterized in that it still comprises the cream. In other cases, e.g. in products which are heat treated after addition of the cream, the cream might be subjected to major alterations in its structure. The present invention is to be understood to encompass all foodstuffs and cosmetic preparations during whose manufacturing process a cream according to the invention was added, regardless of whether the initial structure of the cream is altered in the final product or not.

In the following section, the present invention is further illustrated by examples, which are, however, not intended to limit the scope of the invention as defined in the appended claims.

EXAMPLES

1. Manufacture of Alternan

Plasmid pAI-B-AlSu Q29 was transformed in E. coli DHSa. Vector pAI-B-AlSu contains the full-length coding sequence of alternansucrase derived from Leuconostoc mesenteroides strain NRRL B-1355 (cf. WO 00/47727), lacking the N-terminal 39 amino acids from the signal peptide, fused to an octapeptide strep-tag at the C-terminal end. The strep-tag is linked to the protein through a dipeptide linker. Expression of alternansucrase is under the transcriptional control of the tetA promoter/operator and repressor. The tetA promoter is tightly regulated by the tet repressor which is encoded on the same plasmid and is constitutively expressed from the β-lactamase promoter. In this way, expression of alternansucrase is stringently repressed until efficient chemical induction by tetracycline or anhydrotetracycline, AHT.

The cells were pre-cultured in mineralmedium (Horn et al., 1996) with 100 μg/ml Ampicillin and 10% LB medium. Mineralmedium, without LB, was inoculated with this pre-culture. The cells were grown at 37° C., induced with Anhydrotetracyclin (AHT) (0.2 mg/L), and grown further at 25° C. The cells were harvested, resuspended in [10 mM MOPS pH 7,6; 2,5 mM CaCl2 and 0.05% Trition X-100] and extracted with a high pressure homogenisator. The cell lysate was centrifuged at 20 000 rpm for 20 minutes at 4° C. The supernatant was filtered over a 0.22 μm filter.

Alternan was produced in a 60 L Biotransformation containing 0.13% Acetic Acid, 100 mM NaAc pH5.3, 20% Sucrose, 1 mM DTT, 1600 ml filtered protein extract (ca. 3900 Units). The reaction mixture was incubated for 60 h. at 37° C. The polymer was precipitated with 60L Technical Ethanol 40 h 4° C., washed 2× with 60 L 60% Technical Ethanol, and lx with 60 L 60% Ethanol Absolute. The product was dried through lyophilization

Reference:

Horn U, Strittmatter W, Krebber A, Knupfer U, Kujau M, Wenderoth R, Muller K, Matzku S, Pluckthun A, Riesenberg D. High volumetric yields of functional dimeric miniantibodies in Escherichia coli, using an optimized expression vector and high-cell-density fermentation under non-limited growth conditions. Appl Microbiol Biotechnol 1996; (46): 524-532.

2. Application Tests

2.1 Mayonnaise

Background:

The objective was to test the alternan as produced in example 1 (hereinafter “altenan”) as a fat replacer in a model mayonnaise type dressing system. A model mayonnaise dressing was developed with the control mayonnaise at 68% oil (the standard of identity for mayonnaise is >65% oil).

Three methods of incorporating the alternan were evaluated:

-   -   1. Making an alternan cream and adding it in the last step after         the oil.     -   2. Dry blending the alternan with dry ingredients and adding it         to the egg mixture.     -   3. Making an alternan cream and adding it to the egg mixture.

After the initial experiments using Test Method 1, it was determined that the level of alternan needed to be higher.

Fat replacement levels of 25% to 50% were tested.

Control Formula and Method:

Ingredients % Egg Mixture Egg Yolks 11.15 Water 9.17 Vinegar (Distilled White, 5% Acidity) 8.78 Sugar 1.30 Salt 0.81 Mustard Powder 0.64 Mayonnaise Egg Mixture (from above) 31.85 Salad Oil (Soybean) 68.15

-   -   The egg yolks, water, vinegar (5% acidity), sugar, salt and dry         mustard were mixed together and heated slowly until just         beginning to boil.     -   The egg mixture was allowed to cool slightly and then soybean         oil was sheared into the mixture using a Silverson High Shear         mixer fitted with a Square Hole High Shear screen to form an         emulsion.     -   The mayonnaise was refrigerated overnight before measuring the         viscosity (sample temperatures ranged from 3.3 to 6.7° C.).         Viscosity was measured with a Brookfield RV at Speed 10 with         Spindle #5 or 6, the reading was recorded after 1 minute.

Test Method 1:

Ingredients Alternan Cream % Water 80.91 alternan 10.0 Salad Oil (Soybean) 9.09 Mayonnaise 50% Fat Repl. 25% Fat Repl. Egg Mixture (as described in Control) 31.85 31.85 Salad Oil (Soybean) 34.15 50.15 Alternan Cream 34.00 18.00 Final % of alternan in 50% Fat Replaced Mayonnaise = 3.4% Final % of alternan in 25% Fat Replaced Mayonnaise = 1.8%

-   -   A cream was prepared with water, 10% of the alternan and 9.09%         Soybean Oil (using the Silverson High Shear mixer).     -   The mayonnaise was prepared as described in the Control method         but after the addition of the Soybean Oil, the alternan Cream         was sheared in.

“Blank”

-   -   A sample with a 25% reduction in oil was prepared by simply         increasing the water (no alternan cream was added).

Results and Observations, Test Method 1:

The formula and process for preparing the alternan cream worked well. The cream was white with a medium viscosity. However, the dressings prepared with the alteman cream were much thinner than the control. The test product with a 25% reduction in oil had some viscosity and was similar to a pourable style dressing. The Test product with a 50% reduction in oil was very thin. The level of alternan may have been too low.

Viscosity Sample pH Aw (centapoise)* Observations Control 4.09 0.964 33,000 Thick, forms “peaks” Test with alternan, 4.27 0.984 5,500 Thin 50% oil replacement Test with alternan, 4.24 0.978 9,500 Some thickness/ 25% replacement slight gel Test, Blank with 4.27 0.982 6,200 Some thickness/ 25% replacement slight gel

Test Method 2:

The alternan powder was dry blended with the sugar, salt and mustard. The Control procedure was then followed. A 50% reduction in fat was tested using 10% alternan and 5% alteman. The amount of water in the formula was adjusted to compensate for the reduction in oil and the addition of the alternan to balance the formula to 100%.

Results and Observations, Test Method 2:

The 10% level of alternan was too high for this application. It was very difficult to incorporate the alternan into the egg mixture and a foodstuff processor to mix the mayonnaise had to be used. The resulting product was more like a shortening or spread than a spoonable dressing. The 5% level worked well with the method as described and the resulting product was similar to Control with respect to appearance, texture and viscosity.

Viscosity Sample pH (centapoise) Observations Control (same as described 4.09 33,000 Thick, forms “peaks” in Method 1) 50% Reduced Fat with 10% 1,060,000*   Similar to shortening alternan or butter, fairly stiff and slightly gel like with a smooth/fatty feel. Oil droplets are very small and uniform (microscopic examination). 50% Reduced Fat with 5% 39,700 Thick, similar to alternan, Sample 1 control mayonnaise. Has a smooth, fatty feel when rubbed between the fingers. Very small and uniform oil droplets (microscopic examination). 50% Reduced Fat with 5% 4.14 22,000 Thinner than the alternan, Sample 2 control but still like a mayonnaise. Very fine and uniform oil droplets (microscopic examination). *Since the sample was so thick the viscosity measurement procedure was modified: the Brookfield with Heliopath Spindle F was used at speed 5.

Test Method 3

Ingredients % Egg Mixture Egg Yolks 11.15 Water 0 Vinegar (Distilled White, 5% Acidity) 8.78 Alternan Cream* 48.25 Sugar 1.30 Salt 0.81 Mustard Powder 0.64 *Alternan Cream Water 79.30 alternan 10.36 Salad Oil (Soybean) 10.34 Mayonnaise Egg Mixture 70.93 Salad Oil (Soybean) 29.07 Final level of alternan = 5%

Alteman Cream

-   -   Add the alternan into the water slowly while shearing with the         Silverson mixer.     -   After all the alternan is incorporated, add the oil slowly until         well blended.     -   Follow the control procedure for the egg mixture but whisk the         alternan Cream in last.     -   Follow the control procedure for preparing the mayonnaise.

Results and Observations, Test Method 3:

The method worked well to prepare the alternan cream and it blended into the egg mixture easily. The egg mixture was thick but flowable. The finished mayonnaise was thicker than the control.

Viscosity Sample pH (centapoise) Observations Control, 2^(nd) Sample 4.16 35,700 Thick, typical mayonnaise. Oil droplets are mostly very small with a few larger ones (microscopic examination). 50% Reduced Fat with 5% 4.17 68,000 Thicker than the alternan control but still like a mayonnaise. Oil droplets look the same as control (microscopic examination).

Discussion and Conclusions: The alternan works well to replace 50% of the fat in a spoonable dressing/mayonnaise system at a 5.0% level. A lower level (3.4%) did not add enough viscosity to be comparable to the control. At the 5.0% level, a fairly large variation in the sample viscosity was found when producing a replicate sample on a different date and when using an alternate method. When the alternan was dry blended with other dry ingredients and then added to the egg mixture, resulting viscosities were 39,700 cps and 22,000 cps. However, when the alternan was first made into a cream and then incorporated into the egg mixture the resulting product viscosity was 68,000 cps.

2.2 Pourable Dressing

Background:

The objective was to test the carbohydrate as a fat replacer in a model pourable dressing system. We initially tested a 50% reduction in fat and tested several oil types. We later tested a further reduction in fat with one oil type. The reference material selected was Avicel (FMC BioPolymers, microcrystalline cellulose and carboxymethyl cellulose), a carbohydrate commonly used in low and no fat salad dressings.

Methods:

Control Test with alternan Reference Ingredients % % % Water 21.35 38.85 42.6 Xanthan Gum 0.25 0.25 0.25 Propylene Glycol Alginate 0.15 0.15 0.15 alternan 0.00 5.00 0.00 Avicel SD 3410 0.00 0.00 1.25 Salt 1.75 1.75 1.75 High Fructose Corn Syrup 21.0 21.0 21.0 Vinegar (120 grain) 10.5 10.5 10.5 Vegetable Oil 45.00 22.5 22.5

-   -   Dry blend the gums (and alternan or Avicel) and add them to the         water while shearing with the Silverson High Shear mixer.     -   Shear for 5 minutes. Add the salt, HFCS and vinegar and mix.     -   Slowly add the oil while shearing and shear for an additional         1.5 minutes after all the oil is added.

Note: a “blank” dressing was also prepared with 50% reduced oil but no alternan or avicel in order to determine the viscosity contribution of the xanthan and PGA gums. The Avicel was tested at 0.5, 1.0 and 1.25% but the highest level was closest in viscosity to the control (still lower). An additional sample was also prepared as a reference in which the two gums were increased to levels recommended for a dressing of approximately 20% oil (0.40% xanthan gum and 0.25% PGA).

Results and Observations, Oil Comparison:

The following table summarizes the results for all the oils tested

Cold* Ambient** Viscosity Viscosity Observations and Microscopic Variable (cps) (cps) pH Aroma (Ambient) Evaluation Control with 45% 15,200 13,200 3.51 Oily, slightly painty, Pale yellow/beige color, no Soybean Oil, pungent vinegar. separation. Fairly uniform oil droplets, range about 20 to 30 microns. Control with 45% 14,000 11,040 3.50 Oily, slightly painty, Pale yellow/beige color, no Soybean Oil, pungent vinegar. separation. Fairly uniform oil droplets, range about 20 to 30 microns. Test with 22.5% 12,880 9,200 3.54 Mild, slightly oily, slight Whiter/brighter than control, no Soybean Oil, painty, low pungent. separation. Small to medium oil droplets in the range of 2 to 20 microns. Test with 22.5% 13,400 8,800 3.55 Mild, slightly oily, slight Whiter/brighter than control, no Soybean Oil, painty, low pungent. separation. Fairly uniform oil droplets, most roughly 6 to 10 microns. Test with 22.5% 13,320 9,200 3.54 Very mild, little to no Slightly more beige versus the control, Corn Oil pungency. no separation. Small to medium droplets in the range of 2 to 20 microns. Test with 22.5% 13,680 9,600 3.54 Mild, slightly floral, very Slightly darker/more tan than control, Canola Oil slight pungent. no separation. Small to medium oil (Rapeseed) droplets in the range of 2 to 20 microns. Test with 22.5% 12,600 9,200 3.55 Mild, slightly nutty, very low Slightly whiter than the control, no Sunflower pungent. separation. Small to medium oil Oil droplets in the range of 2 to 20 microns. Test with 22.5% 14,280 9,200 3.57 Fruity oil, no pungency. Greenish/yellow color compared to Extra Virgin the control, no separation. Small to Olive Oil medium oil droplets in the range of 2 to 20 microns. Test “Blank” 3,440 2,720 3.42 Oily, slightly painty, mild Slightly whiter/brighter than control, with 22.5% pungency. no separation. Oil droplets not Soybean Oil uniform, in the range of 5 to >40 microns. Reference with 7,440 6,400 3.54 Oily, slightly painty, mild Slightly whiter than the control, no 22.5% pungency. separation. Small to medium oil Soybean Oil, droplets in the range of 2 to 20 Increased microns. Xanthan and PGA Gums Reference with 7,720 6,320 3.67 Moderate oil, slightly Slightly more white versus the control, 22.5% painty, some pungency. no separation. Oil droplets in the Soybean Oil, range of 2 to 30 microns. 1.25% Avicel SD 3410 Control with 45% 11,200 6,320 3.49 Very fishy. Some separation noted, 3-5 mm of Tuna Oil clear liquid on top, the color is more opaque on the bottom. Oil droplets are not uniform. Size range is about 2 to 100 microns. Test with 22.5% 15,880 8,600 3.53 Somewhat fishy, not No separation noted but a slight oil Tuna Oil pungent. “slick” on the top. Oil droplets are more uniform and smaller versus the control. Most are about 2 to 30 microns, some larger. Reference with 9,800 6,280 3.62 Somewhat fishy, not No separation noted but a slight oil 22.5% Tuna pungent. “slick” on the top. Oil droplets are in Oil, 1.25% the range about 2 to 50 microns, Avicel SD 3410 some larger, many elongated versus round. *Cold Viscosity: Measured on samples immediately after removing from the refrigerator (range 3.3 to 5.6° C.). **Ambient Viscosity: Measured on samples at room temperature (range 20 to 20.6° C.).

Stability Study

Several of the samples were tested for stability by putting them at 37.8° C. and evaluating them at 3 and 7 days. Note that the Tuna Oil samples were not produced in time to include them in the study.

The following table summarizes the results of the stability test:

Results: Day 3 Day 7 “Initial” Viscosity Day 3 Viscosity Day 7 Viscosity Equilibrated Viscosity Equilibrated Viscosity Sample sample, not Sample Day 3 sample, not Sample Day 7 Sample shaken shaken shaken Observations shaken, shaken Observations Control 12,960 13,480 12,080 No separation, 12,880 10,520 Same as Day 3 color similar, oil droplets in the range of 20 to 40 microns. Test with 8,880 12,680 9,360 No separation, 12,680 9,200 About ¼″ of Soybean color similar, oil clear liquid noted Oil droplets in the on the bottom of range of 2 to 20 the jar. Oil droplets microns. look the same as Day 3. Test with 9,000 12,280 9,200 No separation, 11,760 8,760 About ¼″ of Corn Oil color similar, oil clear liquid noted droplets in the on the bottom of range of 2 to 20 the jar. Oil droplets microns. look the same as Day 3. Test with 9,520 13,040 9,680 No separation, 12,080 9,000 About ¼″ of Canola Oil color similar, oil clear liquid noted droplets in the on the bottom of range of 2 to 20 the jar. Oil droplets microns. look the same as Day 3. Test with 9,320 13,080 9,400 No separation, 12,160 9,200 About ¼″ of Sunflower color similar, oil clear liquid noted Oil droplets in the on the bottom of range of 2 to 20 the jar. Oil droplets microns. look the same as Day 3. Test with 9,480 12,800 9,920 No separation, 13,000 9,440 About ¼″ of Olive Oil color slightly clear liquid noted browner, oil on the bottom of droplets in the the jar. Oil droplets range of 2 to 20 look the same as microns. Day 3. Reference 6,560 7,080 6,000 No separation, 6,480 5,600 Same as Day 3 with color similar, oil (no separation Increased droplets in the noted). Gums range of 2 to 20 microns. Reference 6,600 8,480 6,800 No separation, 8,200 6,800 No separation with 1.25% color similar, oil noted. Most oil Avicel droplets in the droplets in the range of 2 to 20 range of 2 to 20 microns. can see microns, some up “fibers” under to 40 microns. microscope.

Further Reduced Fat Dressing

In order to determine if the alternan would work in further reducing the fat in the salad dressing, samples with 10% fat (reduced from 45%) were prepared. At 10% fat, the salad dressing would qualify for a low fat claim which is ≦3.0 grams of fat per serving, serving size is 30 grams. Two higher levels of alternan were tested: 6.5% and 7.75% (7.75% was tested first and the sample was thicker than the control). The samples were prepared as described previously. After preparation, the samples were refrigerated overnight and measured for viscosity. The samples were then allowed to come to ambient temperature and viscosity was checked again. We then put the samples at 37.8° C. to test stability and evaluated at 3 and 7 days

A table summarizing the results follows:

Day 3 Day 3 Day 7 Day 7 Sample Refrig. Ambient Viscosity Viscosity Viscosity Viscosity Description pH Viscosity Viscosity Equilibrated Shaken Equilibrated Shaken Observations 10% SBO, 3.58 32,320 26,400 33,000 33,300 41,500 38,200 No separation at 7.75% 3 days, a slight alternan “crack” near top at 7 days. Oil droplets in the range of 2 to 20 microns, most are <10 microns. 10% SBO, 3.58 10,520 8,120 12,800 8,640 13,080 10,200 No separation at 6.5% 3 days, a slight alternan amount of clear liquid at the bottom of the jar (about 0.25 cm) at 7 days. Oil droplets in the range of 2 to 20 microns, most are <10 microns.

Discussion:

The alternan works well to partially replace the fat in a reduced fat salad dressing. The alternan acts to build back the viscosity which is lost when the oil is decreased. The appearance and aroma of the test dressings was fairly comparable to the Control and the alternan may act to mask some of the pungent aroma. There didn't seem to be an interaction between the alternan and the oil type. The thickness of the dressing seemed to be related to the iodine value of the oil used, the lower the iodine value, the less double bonds, the thicker it would tend to be at refrigerated conditions. There did seem to be a slight advantage to using the alternan in a fish oil containing dressing. The control dressing separated and the oil droplets were larger versus the test dressing with alternan which did not separate and had smaller oil droplets. The alteman containing dressings were able to stand up to some abuse (3 days at 37.8° C.) some separation after 7 days at 37.8° C. was seen. Even though there was some separation, the viscosities remained quite stable. We did note some shear thinning in the Test samples. When samples were allowed to “equilibrate” in a beaker the viscosity was higher versus if the sample was shaken just before the viscosity measurement. We didn't find this to the same extent with the Control dressing. It was possible to reduce the fat in the model dressing to 10% or “low fat” by increasing the alternan to 6.5%. The resulting dressing had similar characteristics to the Control with respect to viscosity and appearance. There was some slight separation after being stored at 37.8° C. for 7 days but the viscosity remained quite consistent.

2.3 Bakery Goods

Background:

The objective was to test the alternan as a fat replacer in bakery goods. Two model systems (cookie and cake) were tested. For cookie the fat replacement was at 30% and for cake at 30% and 50%.

Sugar Snap Cookie

Formula and Methods

Ingredient Full fat (g) % Test (g) % Wheat flour standard 150.0 45.4 150.0 45.4 Water 25.5 7.7 35.8 10.8 Soda 1.2 0.4 1.2 0.4 Milk powder 7.5 2.3 7.5 2.3 Baking powder 2.3 0.7 2.3 0.7 Salt 1.5 0.5 1.5 0.5 Shortening 67.5 20.4 47.3 14.3 Sugar 75.0 22.7 75.0 22.7 Alternan — 10.0 3.0 sum 330.5 100.0 330.5 100.0 Fat % 21.0 15.0 Fat reduction % — 30.0

Method 1:

a) Full fat cookie (control)

-   -   All solids (flour, sugar, soda, baking powder, salt, milk         powder) were mixed in farinograph for 30 sec     -   Shortening added and mixed for 1 min and 30 sec     -   Water added and mixed for 2 min and 30 sec     -   Dough rolled (height 1 cm) and 4 cookies cut with a cutter (76.2         mm)     -   Cookies placed on baking plan and baked at 193° C. for 10 min

b) Alternan reduced fat cookie

-   -   As described in the control except that alternan powder was         mixed with solids

Method 2:

a) Alternan reduced fat cookie

-   -   Making Alternan gel 22% (Alteman powder added to water and         sheared with Ultra Turax at 11600 rpm for 5 min)     -   All solids (flour, sugar, soda, baking powder, salt, milk         powder) were mixed in farinograph for 30 sec     -   Shortening added and mixed for 1 min and 30 sec     -   Alternan gel added and mixed for 2 min and 30 sec     -   Dough rolled (height 1 cm) and 4 cookies cut with a cutter (76.2         mm)     -   Cookies placed on baking plan and baked at 193° C. for 10 min

Results and Observations:

Parameter Full Fat Test 1 Test 2 Dough weight g 326.6 326.8 324.9 Baked dough g 227.6 241.5 242.9 Dough appearance regular regular regular-firmly Cookie weight g 211.7 223.4 224.1 Cookie diameter cm 8.3 7.7 7.9 Height of all cookies cm 5.0 5.3 5.4 Cookie volume cm³ 270.4 246.7 264.6 Baking loss % 7.0 7.5 7.7 Spread ratio 6.6 5.8 5.9 Shape round slightly oval round Browning normal normal normal Surface smooth coarse coarse Hardness of cookie fresh kg 3.5 5.6 7.1 week kg 6.1 6.2 8.6 Crumb moisture fresh % 9.8 16.1 16.4 week % 9.8 12.8 11.3 Test1 = Cookies with Alternan by method 1 Test2 = Cookies with Alternan by method 2

Cake (Sand Cake)

Formulas and Methods

Full fat Test Test Test Test Test Ingredients % 1% 2% 3% 4% 5% Wheat flour 14 14 14 14 14 14 standard Potato starch 14 14 14 14 14 14 Shortening 25 17.5 12.5 12.5 12.5 12.5 Sugar 21 21 21 21 21 21 Vanilla sugar 1.4 1.4 1.4 1.4 1.4 1.4 Whole egg 24 24 24 24 24 24 Salt 0.1 0.1 0.1 0.1 0.1 0.1 Baking powder 0.5 0.5 0.5 0.5 0.5 0.5 Alternan — 1.5 1.5 3 3 6 Water — 6 11 9.5 9.5 6.5 Sum 100 100 100 100 100 100 Fat % 28 20.5 15.5 15.5 15.5 15.5 Fat 27 45 45 45 45 reduction %

Method 1:

a) Full fat cake (control)

-   -   The solids (flour, salt, starch and baking powder were blended         (mixture)     -   The shortening, sugar and vanilla sugar were mixed in a kitchen         machine for 1 min     -   The whisked eggs then added and mixed for 1 min     -   The mixture was gradually added and mixed for 2 min     -   120 g of dough was placed in a baking pan and baked for 40 min         at 160° C.

b) Alternan reduced fat cake (Tests 1 to 3 and Test 5)

-   -   As described in the control, except that alternan powder was         mixed with solids

Method 2:

a) Alternan reduced fat cookie (Test 4)

-   -   Making Alternan gel 24% (Alternan powder added to water and         sheared with Ultra Turax at 11600 rpm for 5min)     -   The solids (flour, salt, starch and baking powder were blended         (mixture)     -   The Alternan gel, shortening, sugar and vanilla sugar were mixed         in a kitchen machine for 1 min     -   The whisked eggs then added and mixed for 1 min     -   The mixture was gradually added and mixed for 2 min     -   120 g of dough was placed in a baking pan and baked for 40 min         at 160° C.

Results and Observations:

Parameter Full fat Test 1 Test 2 Test 3 Test 4 Test 5 Dough consistency creamy/firm less firm less firm less firm less firm less firm Dough moisture % 21.7 27.7 31.3 30.8 30.4 29 Cake weight g 106.9 104.4 102.7 103.2 102.9 103.6 Baking loss % 10.9 13 14.4 14 14.25 13.7 Cake volume ml 575 500 420 420 410 515 Cake moisture % % 12.1 16.9 19.7 19.5 18.8 17.8 Crumb moisture % fresh 21.1 27.3 33.6 31.4 32.2 28.4 week 12 18.1 22.5 22.2 23.2 19.1 Cake height mm 50.2 47.6 44.1 46.4 44.9 48.1 Crumb firmness g fresh 337 296 313 285.4 319.3 213.9 week 1282 1162 1255 1170 1028 1285 Cake browning golden brown golden brown golden brown golden brown golden brown golden brown Crumb appearance succulent/dry succulent/less succulent/less succulent/slightly irregularly succulent/less (normal) dry dry moist baked dry 

1-6. (canceled)
 7. A homogeneous composition comprising alternan, at least one fat or oil, and water. 8-9. (canceled)
 10. The homogeneous composition of claim 7, comprising, 5-15 weight-% alternan, 70-90 weight-% water, and 5-15 weight-% fat and/or oil.
 11. The homogeneous composition, comprising alternan and milk.
 12. The homogeneous composition of claim 7, comprising 5-15 weight-% alternan, and 85-95 weight-% milk.
 13. A method for preparing the homogeneous composition of claim 7, comprising a) adding alternan and water into a vessel b) applying shear to obtain a homogeneous composition, and optionally c) adding oil and/or fat and applying shear to obtain a homogeneous composition.
 14. A method for preparing the homogeneous composition claim 11, comprising a) adding alternan and milk into a vessel, and b) applying shear to obtain a homogeneous composition.
 15. A method for manufacturing foodstuffs or cosmetic preparations comprising adding the homogeneous composition of claim 7, wherein the composition is added as a fat or oil replacer in foodstuffs.
 16. (canceled)
 17. The method of claim 15, wherein the foodstuff is a dairy product, yoghurt, ice cream, milk-based soft ice, milk-based garnish, pudding, cream, whipped cream, chocolate cream, butter cream, crème fraiche, curd, milkshake, egg custard, cheese, nutrition bar, energy bar, breakfast bar, confectionery, bakery product, cracker, cookie, biscuit, cereal chip, snack product, infant and baby foodstuff, bread, croissant, spread, sugar-free biscuit and chocolate, calcium chew, meat product, sausage, mayonnaise, dressing, nut butter, deep-frozen meal, sauce, gravy, soup, shortening, canned food and or ready-to-serve meal.
 18. A foodstuff comprising (a) a foodstuff and (b) an added homogeneous composition including: (i) alternan and water; or (ii) alternan and milk, wherein the homogeneous composition is added as a constituent.
 19. A cosmetic preparation comprising (a) a cosmetic and (b) an added homogeneous composition including: (i) alternan and water; or (ii) alternan and milk, is added as a constituent.
 20. The foodstuff of claim 18, wherein the homogeneous composition comprises: 5-15 weight-% alternan, and 85-95 weight-% water.
 22. The cosmetic preparation of claim 19, wherein the homogeneous composition comprises: 5-15 weight-% alternan, and 85-95 weight-% water.
 23. The cosmetic preparation of claim 19, wherein the homogeneous composition comprises: 5-15 weight-% alternan, and 85-95 weight-% milk.
 24. A method of emulsifying a product comprising adding a alternan.
 25. The method according to claim 24, wherein the alternan has a weight average molecular weight (Mw) in the range of 10 000 000 g/mol to 60 000 000 g/mol, as determined with GPC MALLS.
 26. The method according to claim 24, wherein the alternan has a weight average molecular weight (Mw) in the range of 33 000 000 g/mol to 60 000 000 g/mol, as determined with GPC MALLS.
 27. The method according to claim 24, wherein the alternan has a weight average molecular weight (Mw) in the range of 12 000 000 g/mol to 30 000 000 g/mol, as determined with GPC MALLS.
 28. The method according to claim 24, wherein the product is foodstuffs.
 29. The method according to claim 28, wherein the foodstuff is a dairy product, yoghurt, ice cream, milk-based soft ice, milk-based garnish, pudding, cream, whipped cream, chocolate cream, butter cream, crème fraiche, curd, milkshakes, egg custard, cheese, nutrition bar, energy bar, breakfast bar, confectionery, bakery product, cracker, cookie, biscuit, cereal chip, snack product, infant and baby foodstuff, bread, croissant, spread, sugar-free biscuit and chocolate, calcium chew, meat product, sausage, mayonnaise, dressing, nut butter, deep-frozen meal, sauce, gravy, soup, shortening, canned food or ready-to-serve meal.
 30. The method of claim 29, wherein the cheese is a cream cheese, soft cheese, sliced cheese, or hard cheese.
 31. The method according claim 24, wherein the product is a cosmetic preparation.
 32. The method according to claim 31, wherein the cosmetic preparation is a deodorant, antiperspirant, antiperspirant/deodorant, shaving product, skin cream and lotion, moisturizer, toner, bath product, cleansing product, hair care product, nail cream and lotion, cuticle softener, protective cream, and other personal care formulations.
 33. The method of claim 32, wherein the hair care product is a shampoo, conditioner, mousse, or styling gel.
 34. The method of claim 32, wherein the protective cream is a sunscreen or anti-aging product. 